Wu Jieyun, Li Dongmei, Balan Rebijith K, George Sherly, Peacock Lora, Pal Chandan
Plant Health and Environment Laboratory, Biosecurity New Zealand, Ministry for Primary Industries, P.O. Box 2095, Auckland 1140, New Zealand.
School of Biological Sciences, The University of Auckland, 23 Symonds Street, Auckland 1010, New Zealand.
Insects. 2025 May 27;16(6):564. doi: 10.3390/insects16060564.
Biting midges, spp. (Diptera: Ceratopogonidae), are significant vectors capable of transmitting arboviruses, such as bluetongue virus, to livestock. New Zealand is free of , and a national surveillance programme is in place for the early detection of an incursion. Traditionally, insect trap samples from the surveillance programme are analyzed using morphology-based diagnostics under microscopes, which is time-consuming and relies on specialized taxonomic expertise. Here, we assessed the effectiveness of DNA metabarcoding using insect bulk samples and environmental DNA (eDNA) from liquid samples collected in surveillance traps. Two Cytochrome oxidase I (COI) barcoding primer sets were employed to study biodiversity and detect exotic species. The results indicated that DNA metabarcoding with homogenized insect bulk samples had a higher overall detection accuracy rate (over 81% for both primer pairs) compared to ethanol fluid-derived eDNA samples from traps (68.42% and 55.26% for the primer sets LCO1490/HCO2198 and mlCOIintF/jgHCO2198, respectively) based on congruence with morphological identification. Detection failures were likely due to eDNA extraction issues or low target species abundance. Both approaches showed similar insect community composition and diversity in the surveillance trap samples, suggesting the potential of DNA metabarcoding for biosecurity surveillance and biodiversity assessments. Overall, DNA metabarcoding using bulk insect samples could enhance the efficiency of surveillance, reducing workload and screening time.
蠓科（双翅目：蠓科）昆虫是重要的病媒，能够将虫媒病毒，如蓝舌病毒传播给家畜。新西兰没有[具体蠓种名称未给出]，并且已实施国家监测计划以尽早发现其入侵情况。传统上，监测计划中的昆虫诱捕样本是在显微镜下使用基于形态学的诊断方法进行分析的，这既耗时又依赖专业的分类学专业知识。在此，我们评估了使用昆虫混合样本和监测诱捕器收集的液体样本中的环境DNA（eDNA）进行DNA宏条形码分析的有效性。使用了两套细胞色素氧化酶I（COI）条形码引物来研究生物多样性并检测外来物种。结果表明，与基于形态学鉴定的一致性相比，使用匀浆昆虫混合样本进行DNA宏条形码分析的总体检测准确率更高（两种引物对均超过81%），而诱捕器中乙醇液体衍生的eDNA样本的检测准确率分别为68.42%和55.26%（引物组LCO1490/HCO2198和mlCOIintF/jgHCO2198）。检测失败可能是由于eDNA提取问题或目标物种丰度较低。两种方法在监测诱捕器样本中显示出相似的昆虫群落组成和多样性，表明DNA宏条形码分析在生物安全监测和生物多样性评估方面具有潜力。总体而言，使用昆虫混合样本进行DNA宏条形码分析可以提高[具体蠓种名称未给出]监测的效率，减少工作量和筛查时间。
